In any suspension culture where all cells have similar density, why do the cells float in different layers inside the media? Is it necessary to keep a flask with these cultures in a vertical position (inside the CO2 incubator)?
Dear Deepa,
in my experience, cells in suspension will usually sink to the bottom of your culture dish if they are kept in static incubators (or even just in a centrifuge tube if kept there for a while). If they are true suspension cells, they won't adhere to the plastic, but they will sit there until disturbed. You can observe this easily when you seed for example splenocytes into 96well round bottom plates. The cells will all sink down to the bottom and form a 'pellet', the medium will be clear. Only when you have just seeded your cells or take your dishes out from the incubator you can't avoid shaking them as Omar suggests and they will start swirling around.
However, if you have a very bad contamination in your cultures (especially yeast...) they might appear to be everywhere and your medium may look turbid even before shaking or moving.
Susanne
It could be because the brownian movement or because you are shaking the media?
Dear Deepa,
in my experience, cells in suspension will usually sink to the bottom of your culture dish if they are kept in static incubators (or even just in a centrifuge tube if kept there for a while). If they are true suspension cells, they won't adhere to the plastic, but they will sit there until disturbed. You can observe this easily when you seed for example splenocytes into 96well round bottom plates. The cells will all sink down to the bottom and form a 'pellet', the medium will be clear. Only when you have just seeded your cells or take your dishes out from the incubator you can't avoid shaking them as Omar suggests and they will start swirling around.
However, if you have a very bad contamination in your cultures (especially yeast...) they might appear to be everywhere and your medium may look turbid even before shaking or moving.
Susanne
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